Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Rheumatology Research Center, Tehran University of Medical Sciences, Tehran, Iran.
Inflammopharmacology. 2024 Oct;32(5):3181-3193. doi: 10.1007/s10787-024-01536-6. Epub 2024 Aug 21.
Systemic sclerosis (SSc) is a connective tissue disorder characterized by excessive fibrosis, where activated fibroblasts play a pivotal role in disease progression. This study aimed to investigate the potential of Talabostat, a small molecule inhibitor of dipeptidyl peptidases, in alleviating fibrosis and inflammation associated with SSc pathogenesis.
Dermal fibroblasts were obtained from skin biopsies of ten diffuse cutaneous SSc patients and healthy controls. These fibroblasts were subjected to treatment with either TGF-β alone or in combination with Talabostat. Immunofluorescence staining was conducted to evaluate FAPα and α-SMA protein levels. The expression of activated fibroblast markers (FAPα and ACAT2), pro-fibrotic (COL1A1 and COL1A2), anti-fibrotic (MMP1, MMP2, and MMP9), and inflammatory (IL-6 and TGFβ1) related genes was measured by quantitative real-time PCR. Talabostat-treated fibroblasts were assessed for their migratory capacity using a scratch assay and for their viability through MTT assay and Annexin V staining.
The basal expression of COL1A1 and TGFβ1 was notably higher in healthy subjects, while MMP1 expression showed a significant increase in SSc patients. Furthermore, TGF-β stimulation led to upregulation of activated fibroblast markers, pro-fibrotic, and inflammatory-related genes in SSc-derived fibroblasts, which were attenuated upon Talabostat treatment. Interestingly, Talabostat treatment resulted in an upregulation of MMP9 expression. Moreover, Talabostat exhibited a concentration-dependent inhibition of activated fibroblast viability in both healthy and SSc fibroblasts, and suppressed fibroblast migration specifically in SSc patients.
In summary, Talabostat modulates fibrotic genes in SSc, thereby inhibiting myofibroblast differentiation, activation, and migration. These findings suggest promising therapeutic avenues for targeting fibrosis in SSc.
系统性硬化症（SSc）是一种结缔组织疾病，其特征是过度纤维化，其中活化的成纤维细胞在疾病进展中起着关键作用。本研究旨在探讨小分子二肽基肽酶抑制剂 Talabostat 缓解与 SSc 发病机制相关的纤维化和炎症的潜力。
从 10 例弥漫性皮肤 SSc 患者和健康对照者的皮肤活检中获取真皮成纤维细胞。这些成纤维细胞用 TGF-β 单独或与 Talabostat 联合处理。通过免疫荧光染色评估 FAPα 和 α-SMA 蛋白水平。通过定量实时 PCR 测量激活的成纤维细胞标志物（FAPα 和 ACAT2）、促纤维化（COL1A1 和 COL1A2）、抗纤维化（MMP1、MMP2 和 MMP9）和炎症（IL-6 和 TGFβ1）相关基因的表达。通过划痕实验评估 Talabostat 处理的成纤维细胞的迁移能力，通过 MTT 实验和 Annexin V 染色评估成纤维细胞的活力。
健康受试者中 COL1A1 和 TGFβ1 的基础表达明显较高，而 SSc 患者中 MMP1 的表达显著增加。此外，TGF-β 刺激导致 SSc 来源的成纤维细胞中激活的成纤维细胞标志物、促纤维化和炎症相关基因的上调，而 Talabostat 处理则减弱了这些基因的上调。有趣的是，Talabostat 处理导致 MMP9 表达上调。此外，Talabostat 以浓度依赖的方式抑制健康和 SSc 成纤维细胞中激活的成纤维细胞活力，并特异性抑制 SSc 患者的成纤维细胞迁移。
总之，Talabostat 调节 SSc 中的纤维化基因，从而抑制肌成纤维细胞分化、激活和迁移。这些发现为 SSc 中靶向纤维化的治疗提供了有希望的途径。
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